Estrogens

ABSTRACT

Compounds of the formula I ##STR1## a process for their preparation, their use in the treatment of autoimmune disorders as well as new intermediates for their preparation.

This application is a 371 of PCT/SE96/01028 filed Aug. 20, 1996.

FIELD OF THE INVENTION

The present invention relates to novel compounds which are steroidalestrogens, to methods for their preparation, their use andpharmaceutical compositions comprising the novel compounds. The novelcompounds are useful for the treatment of inflammatory and immunologicdisorders, especially for the treatment of autoimmune disorders. Thecompounds according to the present invention are especially preferredfor the treatment of rheumatoid arthritis (RA) and multiple sclerosis(MS).

BACKGROUND AND PRIOR ART

Sex hormones have since long been known to ameliorate arthritic symptomsin chronic arthritis during pregnancy, see for example Hench P. S. "Theameliorating effect of pregnancy on chronic atrophic arthritis,fibrositis, and intermittent hydrathrosis", Mayo Clin. Proc., 13,161-167, 1983. The use of oral contraceptives in patients withrheumatoid arthritis (RA) have proven to decrease the incidence of RA,see Wingrave S. J., Kay C. R. "Reduction in incidence of rheumatoidarthritis associated with oral contraceptives", Lancet, 569-571, 1978;and Vandenbroucke J. P. et al., "Oral contraceptives and rheumatoidarthritis: Further evidence for a preventive effect", Lancet 839-842,1982.

In JP 268575/1990 estradiol derivatives are described, but thesubstituents in 17-position are completely different from thesubstituents in 17-position of the present application. The problemunderlying the invention described in JP 268575/1990 is to findcompounds against osteoporosis, said compounds having an excellent boneresorption inhibiting action without showing side effects such as riskfor genital cancer etc. known in the art for estrogens.

The problem underlying the present invention is to develop novelsteroidal estrogens with high anti-inflammatory and immunosuppressiveeffects, but with low "sex hormonal" activities. The steroidal estrogensknown in the prior art, have the disadvantages that they influencegenital and breast tissues, thereby conferring adverse effects such asendometrial and breast cancers if given in too high amounts.

The problem mentioned above has been solved by developing new steroidalestrogens according to the formula I, as will be described in thefollowing.

SUMMARY OF THE INVENTION

The object of the present invention is to provide novel compounds, whichare steroidal estrogens, and a method for their preparation.

Another object of the present invention is the use of the novelcompounds for the treatment of inflammatory and immunologic diseases,especially for the treatment of autoimmune diseases.

Still another object of the invention is a pharmaceutical compositioncomprising a compound of the invention as active ingredient, optionallyin the presence of a pharmaceutically acceptable carrier.

The novel compounds of the present invention are defined by the generalformula I ##STR2## wherein A is hydrogen, C₂ -C₁₈ alkanoyl, (C₆aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl, (C₆ aryloxycarbonyl, or aprotecting group;

B is hydrogen, methyl, or ethyl;

R is hydrogen, a straight, branched or cyclic C₁ -C₆ alkyl, C₂ -C₁₈alkanoyl, (C₆ aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl, (C₆aryloxy)carbonyl, or a protecting group;

X¹ is hydrogen, methyl, ethyl, or halogen;

X² is hydrogen, methyl, ethyl, or halogen; and

Y is methylene or a single bond;

the compounds

(17E)-16α-Acetoxy-3-methoxy-19-norpregna-1,3,5(10),17(20)-tetraene;

(17E)-16α-hydroxy-3-methoxy-19-norpregna-1,3,5(10),17(20)-tetraene; and

(17E)-16β-hydroxy-3-methoxy-19-norpregna-1,3,5(10),17(20)-tetraene

being excluded.

Within the scope of the invention are also pharmaceutically acceptablesalts of the compounds of the formula I.

Preferred compounds of the invention are compounds of the formula Iwherein

A is hydrogen or C₂ -C₆ alkanoyl;

B is hydrogen or methyl;

R is hydrogen, a straight, branched or cyclic C₁ -C₆ alkyl, C₂ -C₁₈alkanoyl, (C₆ aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl, (C₆aryloxy)carbonyl, or a protecting group;

X¹ is hydrogen, methyl, or fluorine;

X² is hydrogen, methyl, or fluorine; and

Y is methylene or a single bond.

Particularly preferred compounds of the invention are compoundsaccording to the formula I wherein

A is hydrogen or C₂ -C₆ alkanoyl;

B is hydrogen;

R is hydrogen, a straight, branched or cyclic C₁ -C₆ alkyl, C₂ -C₁₉alkanoyl or (C₆ aryl)carbonyl;

X¹ is hydrogen or fluorine;

X² is hydrogen or fluorine; and

Y is a single bond or a methylene group.

Examples of protecting groups are benzyl, THP (tetrahydropyranyl),methoxymethyl, dimethylthexylsilyl, and tert-butyldimethylsilyl. Apreferred protecting group is dimethylthexylsilyl.

The most preferred compound of the invention is3,16α-dihydroxy-17-methylene-estra-1,3,5(10)triene.

The novel steroidal estrogens according to the invention arecharacterized by high anti-inflammatory and immunosuppressive effects,and low "sex hormonal" activities. Thus the novel steroidal estrogenshave low proliferative effects on genital tissues, which reduces thelikelihood of adverse effects such as endometrial cancers.

The novel steroidal estrogens according to the invention are useful forthe treatment of inflammatory and immunologic disorders, especially forthe treatment of autoimmune disorders.

The steroidal estrogens according to the present invention are excellentfor the treatment of rheumatoid arthritis (RA) and multiple sclerosis(MS).

DETAILED DESCRIPTION OF THE INVENTION

Methods of Preparation

Common to all starting materials for the preparation of compounds of theformula I is the presence of a 17-keto group. The introduction of the17-alkylidene group can be achieved by a Wittig-type reaction (see e.g.Krubiner, A. M. et al. J. Org. Chem., 1966, 31, 24) whereby a compoundof the formula II ##STR3## wherein A is hydrogen, C₂ -C₁₈ alkanoyl, (C₆aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl, (C₆ aryloxy)carbonyl, or aprotecting group;

B is hydrogen, methyl, or ethyl;

R is hydrogen, a straight, branched or cyclic C₁ -C₆ alkyl, C₂ -C₁₈alkanoyl, (C₆ aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl, (C₆aryl)oxycarbonyl, or a protecting group; and

Q is (O--A) or hydrogen, wherein O is oxygen and A is as defined above;

the 3-O-position being optionally protected is reacted with aphosphorous ylide or with the salt of a stabilized alkylphosphonate,optionally followed by the reduction of the adduct when a stabilizedalkyl phosphonate is used, giving a compound of the formula III ##STR4##wherein A is hydrogen, C₂ -C₁₈ alkanoyl, (C₆ aryl)carbonyl, C₂ -C₁₉alkoxycarbonyl, (C₆ aryloxycarbonyl, or a protecting group;

B is hydrogen, methyl, or ethyl;

R is hydrogen, a straight, branched or cyclic C₁ -C₆ alkyl, C₂ -C₁₈alkanoyl, (C₆ aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl, (C₆aryloxy)carbonyl, or a protecting group; and

X¹ is hydrogen, methyl, ethyl or halogen; and

X² is hydrogen, methyl, ethyl or halogen.

T is (O--A) or hydrogen, wherein O is oxygen and A is as defined above;

the 3-O-position being optionally protected.

The above given definition that Q and T is (O--A) or hydrogenrespectively, means that the 16-position may be protected orunprotected.

The reaction is preferably carried out in a polar solvent such as DMSO,THF or dimethoxyethane, and the temperature is preferably in the rangeambient temperature to the boiling point of the solvent.

When stabilized alkylphosphonates are used, the substituents X¹ and X²in formula III may be carbonyl moieties, such as an ester or ketone,which can be reduced to an alcohol, and further reduced to an alkylgroup.

The 16-OA functionality may be present in the starting material orintroduced at a later stage. If not present in the starting material,the 16-OA functionality is introduced via an oxidation such as a SeO₂-oxidation (Sharpless, K. B. et al. Aldrichimica Acta, 1979, 12, 63),whereby a compound of the formula IV ##STR5## wherein B is hydrogen,methyl, or ethyl;

R is hydrogen, a straight, branched or cyclic C₁ -C₆ alkyl, C₂ -C₁₈alkanoyl, (C₆ aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl, (C₆aryloxy)carbonyl, or a protecting group; and

Each of X¹ and X² is independently hydrogen, methyl, or ethyl;

is subjected to a SeO₂ -oxidation, giving the 16α-OH compound of theformula V selectively (Trost, B. M. et al. J. Am. Chem. Soc., 1978, 100,3435) together with the 16-keto compound of the formula VI ##STR6##wherein B is hydrogen, methyl, or ethyl; R is hydrogen, a straight,branched or cyclic C₁ -C₆ alkyl, C₂ -C₁₈ alkanoyl, (C₆ aryl)carbonyl, C₂-C₁₉ alkoxycarbonyl, (C₆ aryloxy)carbonyl, or a protecting group;

Each of X¹ and X² is independently hydrogen, methyl or ethyl; and

Y is a single bond.

In a compound of the formula VI, X¹ and X² may also each independentlybe selected from a halogen, and Y may also be selected from methylene.

SeO₂ is preferably used in catalytic amounts together withtertbutylhydroperoxide as a co-oxidant in toluene at ambienttemperature.

These first reaction steps can be performed on either 3-O-unprotected(R═H) or protected (R═e.g. R₃ Si, tetrahydropyranyl (THP), alkyl orbenzyl) material. The introduction of the protecting group is achievedby standard methods (Protective groups in organic synthesis, Green, T.W. and Wuts, P. G. M., 2nd ed., Wiley). Thus, the free phenol can beprotected as a dimethylthexylsilyl ether using dimethylthexylsilylchloride as silylating reagent and imidazole as base in the solventdimethylformamide (DMF) at ambient temperature.

The 16-keto compound of the formula VI is further subjected to anucleophile, such as a Grignard reagent in an inert solvent, such as Et₂O or THF, or alternatively reduced, e.g. with NaBH₄ or LiAlH₄, givingthe 16β-hydroxy compound of the formula I wherein Y is a single bond.

The cyclopropane moiety is introduced by reacting a compound of theformula I or VI with a cyclopropanation reagent, whereby the alkenemoiety of the compound of the formula I or VI wherein Y is a singlebond, is reacted with a cyclopropanation reagent, optionally in thepresence of a metal promotor, giving a compound of the formula I or ofthe formula VI (Y═methylene) respectively. One preferredcyclopropanation reaction is the Simmons-Smith reaction, using a1,1-dihalo compound in the presence of activated Zn, preferably inetheral solvents such as dimethoxyethane. The cyclopropanation reactionof choice for the introduction of the cyclopropane moiety will be clearfor one skilled in the art (Advanced Organic Chemistry: reactions,mechanisms and structure, J. March, 4th ed., p 870 ff., Wiley).

The phenolic 3-OH group may be protected, e.g., as a silylether (or asan alkylether, a benzylether, or an acetal, like THP-ether) throughoutthe reaction sequences. Thus, the unprotected 16-OH can then be reactedwith activated ester derivatives, such as ester halides or anhydrides,to give 16-O-monoacylated derivatives.

The 3-O-silyl ether can be cleaved by fluoride ion (e.g. Bu₄ NF(H₂ O)₃in THF) or by acid or base treatment to give the free phenol derivatives(van Look, G., "Silylating Agents", Fluka Chemie, 1988). The3-O-monoacylated derivatives can also be regioselectively prepared, e.g.by acylating the tetrabutylammonium phenolate generated in the Bu₄NF-desilylation step by acylating agents like acid chlorides oranhydrides, or by acylating the 3,16-diol by the method of Illi V. O.,Tetrahedron Lett. 1979, p. 2431 using acid chlorides as acylatingreagents in dioxane, NaOH as base and catalytic amounts oftetrabutylammonium hydrogen sulfate.

EXAMPLES

The invention will now be described in more detail by the followingexamples which are not to be construed as limiting the invention.

In the examples column chromatography separations were performed usingMerck SiO₂ 60 (0.040-0.063 mm) silica gel with heptane-EtOAc mixtures aseluents.

TLC analyses were performed on Merck SiO₂ 60 F254 precoated aluminiumsheets: R_(f) values were measured in heptane-EtOAc eluent mixtures andthe spots were visualized by charring with 10% aqueous H₂ SO₄.

Melting points were determined with a Weitz Wetzlar microscope and areuncorrected.

MS(FAB) spectra were recorded with a VG Analytical Autospec-Qspectrometer. NMR spectra were recorded with a Varian VXR (300 MHz) or aVarian Unity+ (500 MHz).

Dry solvents were prepared by drying p.a. (pro analysis) grade solventsover molecular sieves (4 Å).

General Procedure for 16α-hydroxylation of 17-alkylidenes (Procedure A):

SeO₂ (11 mg, 0.1 mmol) was added to a solution of the 17-alkylidene (1.0mmol) and tert-butylhydroperoxide (0.67 ml, ca 2.0 mmol, ca 3.0 M "phaseseparated" in toluene, Sharpless, K. B. et al. Aldrichimica Acta, 1979,12, 63) in toluene (1.0 ml). The reaction mixture was stirred over nightand thereafter diluted with Et₂ O (50 ml). FeSO₄ (10 ml, 1 M) was addedand after 30 min stirring the organic phase was separated and washedwith brine (2×30 ml). The organic phase was dried over Na₂ SO₄ andconcentrated at reduced pressure. The residue was purified by columnchromatography to give the 17-alkylidene-16α-hydroxy compound (yields ca40-60%) and the 17-alkylidene-16-keto compound (yields ca 20-30%).

General Procedure for 3-O-Desilylation of 3-O-Dimethyl-thexylsilyl EtherProtected 3-hydroxy-estra-1,3,5(10)-trienes (Procedure B):

NBu₄ F·(H₂ O)₃ (1.1 mmol) was added to a solution of the3-dimethyl-thexylsilyl ether protected 3-hydroxy-estra-1,3,5(10)-triene(1.0 mmol) in dry THF (1.0 mL). The reaction mixture was stirred for 3min and thereafter quenched by adding AcOH (1.5 mmol). Concentration atreduced pressure was followed by purification on column chromatography.Alternatively, for larger scale synthesis, a typical work up (dilutionwith Et₂ O, washing with water, drying, and concentration) may precedethe column chromatography.

Silylations were performed according to the Corey procedure (Corey, E.J., Venkateswarlu, A. J. Am. Chem. Soc. 1972, 94, 6190) withdimethyl-thexyl chlorosilane (1.2 mol eq.) as silylating agent andimidazole (2.5 mol eq.) as base in DMF as solvent. Usual work-up(dilution with Et₂ O, washing with water, drying, and concentration)followed by column chromatography provided the products in essentiallyquantitative yields.

Examples 1-2

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene

Prepared from 3-hydroxy-17-methylene-estra-1,3,5(10)-triene (83% fromestrone, Peters, R. H. et al. J. Med. Chem. 1989, 32, 1642) according toProcedure A.

Also prepared from 3,16α-dihydroxy-17-methylene-estra-1,3,5(10)-triene,3-dimethyl-thexylsilyl ether (Example 5) according to Procedure B.

Yield: 356 mg (92%)

R_(f) (2:1)=0.20

mp 245-50° C.

MS(FAB): m/z=284 (M⁺)

¹ H NMR (CDCl₃) δ 0.83 (s, 3H, H-18), 4.53 (s, 1H, phenol), 4.72 (m, 1H,H-16), 4.93 (d, 1H, J=2.1 Hz, ═CH₂), 5.08 (d, 1H, J=1.4 Hz, ═CH₂), 6.57(d, 1H, J=2.8 Hz, H-4), 6.63 (dd, 1H, J=2.8 Hz, 8.3 Hz, H-2), 7.17 (d,1H, J=8.3 Hz, H-1).

Example 3

3-Hydroxy-17-keto-estra-1,3,5(10)-triene, 3-O-dimethylthexylsilyl ether

Prepared from 3-hydroxy-17-keto-estra-1,3,5(10)-triene (estrone) bysilylation using the Corey procedure.

Yield: 29.3 g (94%)

R_(f) (10:1)=0.10

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --SiMe₂ --), 0.91 (s, 3H, H-18), 0.94 (s,6H, thexyl), 0.94 (d, 6H, J=6.6 Hz, thexyl), 6.56 (d, 1H, J=2.7 Hz,H-4), 6.62 (dd, 1H, J=2.7 Hz, 8.3 Hz, H-2), 7.12 (d, 1H, J=8.3 Hz, H-1).

Example 4

3-Hydroxy-17-methylene-estra-1,3,5(10)-triene, 3-O-dimethylthexylsilylether

Prepared from 3-hydroxy-17-methylene-estra-1,3,5(10)-triene (83% fromestrone, Peters, R. H. et al. J. Med. Chem. 1989, 32, 1642) bysilylation using the Corey procedure.

Yield: 22.4 g (99%)

R_(f) (8:1)=0.18

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --SiMe₂ --), 0.82 (s, 3H, H-18), 0.94 (s,6H, thexyl), 0.94 (d, 6H, J=6.6 Hz, thexyl), 4.67 (s, 2H, ═CH₂), 6.55(d, 1H, J=2.8 Hz, H-4), 6.61 (dd, 1H, J=2.8 Hz, 8.3 Hz, H-2), 7.14 (d,1H, J=8.3 Hz, H-1).

Example 5

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether

Prepared from 3-hydroxy-17-methylene-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether according to procedure A.

Yield: 5.07 g (59%)

R_(f) (5:1)=0.29

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --SiMe₂ --), 0.83 (s, 3H, H-18), 0.94 (s,6H, thexyl), 0.94 (d, 6H, J=6.6 Hz, thexyl), 4.71 (m, 1H, H-16), 4.92(d, 1H, J=2.2 Hz, ═CH₂), 5.08 (d, 1H, J=1.7 Hz, ═CH₂), 6.55 (d, 1H,J=2.7 Hz, H-4), 6.61 (dd, 1H, J=2.7 Hz, 8.3 Hz, H-2), 7.13 (d, 1H, J=8.3Hz, H-1).

This reaction also provided the compound of Example 6 below.

Example 6

3-Hydroxy-16-keto-17-methylene-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether

See under Example 5 regarding the synthesis.

Yield: 1.54 g (18%)

R_(f) (5:1)=0.56

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --SiMe₂ --), 0.94 (s, 6H, thexyl), 0.94(d, 6H, J=7.0 Hz, thexyl), 0.99 (s, 3H, H-18), 5.07 (s, 1H, ═CH₂), 5.84(s, 1H, ═CH₂), 6.56 (d, 1H, J=2.5 Hz, H-4), 6.63 (dd, 1H, J=2.5 Hz, 8.3Hz, H-2), 7.13 (d, 1H, J=8.3 Hz, H-1).

Example 7

3,16β-Dihydroxy-17-methylene-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether

CeCl₃ (283 mg, 1.15 mmol) was added to a solution of3-hydroxy-17-methylene-16-keto-estra-1,3,5(10)-triene,3-dimethyl-thexylsilyl ether (488 mg, 1.15 mmol) in dry THF (12 ml)under N₂. The slurry was stirred for 5 min and then LiAlH₄ (44 mg, 1.15mmol) was added. The reaction mixture was stirred at room temperaturefor 15 min, then quenched with 1 M HCl and partitioned in Et₂ O/water.The organic phase was washed with aq. NaHCO₃ (sat.) and brine, driedover Na₂ SO₄ and concentrated at reduced pressure. The residue waspurified by column chromatography (heptane-EtOAc, 5:1) to give the titelcompound (220 mg, 45%) as a white solid.

R_(f) (5:1)=0.16

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --SiMe₂ --), 0.94 (s, 6H, thexyl), 0.94(d, 6H, J=7.0 Hz, thexyl), 1.00 (s, 3H, H-18), 4.55 (m, 1H, H-16), 4.92(s, 1H, ═CH₂), 5.08 (s, 1H, ═CH₂), 6.55 (d, 1H, J=2.7 Hz, H-4), 6.61(dd, 1H, J=2.7 Hz, 8.3 Hz, H-2), 7.13 (d, 1H, J=8.3 Hz, H-1).

Example 8

3,16β-Dihydroxy-17-methylene-estra-1,3,5(10)-triene

Prepared from 3-hydroxy-17-methylene-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether according to procedure B.

Yield: 46 mg (85%)

R_(f) (1:1)=0.42

mp 229-35° C.

MS(FAB): m/z=284 (M⁺)

¹ H NMR (CDCl₃) δ 1.00 (s, 3H, H-18), 4.53 (m, 1H, H-16), 4.94 (d, 1H,J=1.8 Hz, ═CH₂), 5.08 (d, 1H, J=1.8 Hz, ═CH₂), 6.56 (d, 1H, J=2.7 Hz,H-4), 6.64 (dd, 1H, J=2.7 Hz, 8.5 Hz, H-2), 7.16 (d, 1H, J=8.5 Hz, H-1).

Example 9

3,16β-Dihydroxy-16α-methyl-17-methylene-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether

A solution of 3-hydroxy-17-methylene-16-keto-estra-1,3,5(10)-triene,3-dimethylthexylsilyl ether (108 mg, 0.25 mmol) in dry Et₂ O (2 mL) wasadded to MeMgI (1 mmol, 1M in Et₂ O) at 0° C. under N₂. The reactionmixture was stirred at room temperature over night, then quenched with 1M HCl and partitioned in Et₂ O/water. The organic phase was washed withaq. NaHCO₃ (sat.) and brine, dried over Na₂ SO₄ and concentrated atreduced pressure. The residue was purified by column chromatography(heptane-EtOAc, 8:1) to give the title compound (40 mg, 37%) as a whitesolid.

R_(f) (5:1)=0.21

¹ H-NMR (CDCl₃) δ 0.22 (s, 6H, --SiMe₂ --), 0.94 (s, 6H, thexyl), 0.94(d, 6H, J=7.0 Hz, thexyl), 1.03 (s, 3H, H-18), 1.41 (s, 3H, 16-Me), 4.82(s, 1H, ═CH₂), 5.06 (s, 1H, ═CH₂), 6.55 (d, 1H, J=2.7 Hz, H-4), 6.61(dd, 1H, J=2.7 Hz, 8.3 Hz, H-2), 7.11 (d, 1H, J=8.3 Hz, H-1).

Example 10

3,16β-Dihydroxy-16α-methyl-17-methylene-estra-1,3,5(10)-triene

Prepared from3,16β-dihydroxy-16α-methyl-17-methylene-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether according to procedure B.

Yield: 12 mg (93%)

R_(f) (2:1)=0.22

mp 237-39° C.

MS-FAB: m/z=298 (M⁺)

¹ H NMR (CDCl₃) δ 1.03 (s, 3H, H-18), 1.41 (s, 3H, 16-Me), 4.51 (s, 1H,phenol), 4.83 (s, 1H, ═CH₂), 5.07 (s, 1H, ═CH₂), 6.57 (d, 1H, J=2.8 Hz,H-4), 6.63 (dd, 1H, J=2.8 Hz, 8.3 Hz, H-2), 7.16 (d, 1H, J=8.3 Hz, H-1).

Example 11

3,16α-Dihydroxy-17-(1',2'-ethylene)-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether

A slurry of Zn powder (280 mg, 4.28 mmol) in dry dimethoxyethane (DME,4.0 ml) under N₂ was activated by ultra sound treatment for 1.5 h. Asolution of 3,16α-dihydroxy-17-methylene-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether (500 mg, 1.17 mmol) in dry DME (8.0 mL)was added and the temperature was raised to reflux temperature (ca 90°C. in oil bath). CH₂ I₂ (390 ml, 4.83 mmol) was added dropwise and thereaction mixture was stirred at reflux temperature over night. Aftercooling the reaction mixture was partitioned in EtOAc/NH₄ Cl (aq.,sat.). The organic phase was washed with H₂ O, dried over Na₂ SO₄ andconcentrated at reduced pressure. The residue was purified by columnchromatograph (heptane-EtOAc, 8:1) to give the title compound (280 mg,54%).

R_(f) (5:1)=0.28

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --SiMe₂ --), 0.50, 0.72 (2m, 4H,17-ethylene), 0.82 (s, 3H, H-18), 0.94 (s, 6H, thexyl), 0.94 (d, 6H,J=7.0 Hz, thexyl), 4.19 (m, 1H, H-16), 6.56 (d, 1H, J=2.7 Hz, H-4), 6.62(dd, 1H, J=2.7 Hz, 8.3 Hz, H-2), 7.12 (d, 1H, J=8.3 Hz, H-1).

Example 12

3,16α-Dihydroxy-17-(1',2'-ethylene)-estra-1,3,5(10)-triene

Prepared from3,16α-dihydroxy-17-(1',2'-ethylene)-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether according to procedure B.

Yield: 50 mg (74%)

R_(f) (5:1)=0.10

mp 227-32° C.

MS-FAB: m/z=298 (M⁺)

¹ H NMR ((CD₃)₂ SO) δ 0.24-0.40, 0.65 (2m, 4H, 17-ethylene), 0.76 (s,3H, H-18), 4.08 (m, 1H, H-16), 4.35 (d, 1H, J=7.1 Hz, 16-OH), 6.44 (s,1H, H-4), 6.50 (d, 1H, J=8.6 Hz, H-2), 7.02 (d, 1H, J=8.6 Hz, H-1), 9.00(broad s, 1H, 3-OH).

Example 13

3-Hydroxy-17-keto-16α-methyl-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether

Lithium diisopropylamide (2.8 ml, 4.2 mmol, 1.5 M THF-complex inc-hexane) was added to a solution of3-hydroxy-17-keto-estra-1,3,5(10)-triene, 3-O-dimethylthexylsilyl ether(1.50 g, 3.63 mmol) in dry THF (6 ml) under N₂ at 0° C. After stirringfor 1 h the temperature was lowered to -78° C. and Mel (270 μl, 4.3mmol) was added. The reaction mixture was stirred at -78° C. for 5 h,then at ambient temperature over night and was then partitioned inEtOAc/H₂ O. The organic phase was washed with brine, dried over Na₂ SO₄and concentrated at reduced pressure. The residue was purified by columnchromatography (heptane-EtOAc, 20:1) to give the title compound (800 mg,52%).

R_(f) (20:1)=0.23

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --Si(CH₃)₂ --), 0.94 (s, 3H, H-18), 0.94(s, 6H, thexyl), 0.94 (d, 6H, J=6.8 Hz, thexyl), 1.14 (d, 1H, J=7.8 Hz,16-Me), 6.58 (d, 1H, J=2.4 Hz, H-4), 6.62 (dd, 1H, J=2.4 Hz, 8.3 Hz,H-2), 7.13 (d, 1H, J=8.3 Hz, H-1).

Example 14

3-Hydroxy-16α/β-methyl-17-methylene-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether

Potassium tert-butoxide (73 mg, 0.65 mmol) was added to a solution ofmethyltriphenyl-phosphonium bromide (300 mg, 0.84 mmol) in dry DMSO (1.8ml) under N₂. After stirring for 20 min the temperature was raised to75° C. and a solution of3-hydroxy-17-keto-16α-methyl-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether (298 mg, 70 mmol) in dry THF (1.5 ml) wasadded. The reaction mixture was stirred at 75° C. for 1.5 h and was thenpartitioned in Et₂ O/H₂ O. The organic phase was washed with H₂ O, driedover Na₂ SO₄, and concentrated at reduced pressure. The residue waspurified by column chromatography (heptane) to give the title compoundas a ca 1:1 epimeric mixture (85 mg, 28%).

R_(f) (heptane)=0.24

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --Si(CH₃)₂ --), 0.84, 0.94 (2s, 3H,H-18), 0.94 (s, 6H, thexyl), 0.94 (d, 6H, J=6.8 Hz, thexyl), 1.10, 1.19(2d, 3H, J=7.1 Hz, 16-Me), 4.68, 4.73 (2m, 2H, Hz, ═CH₂), 6.56 (d, 1H,J=2.2 Hz, H-4), 6.62 (dd, 1H, J=2.2 Hz, 8.3 Hz, H-2), 7.13 (d, 1H, J=8.3Hz, H-1).

Example 15

3,16α-Dihydroxy-16α,β-methyl-17-methylene-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether

Prepared from3-hydroxy-16α,β-methyl-17-methylene-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether according to procedure A.

Yield: 35 mg (40%)

R_(f) 10:1)=0.10

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --Si(CH₃)₂ --), 0.87 (s, 3H, H-18), 0.94(s, 6H, thexyl), 0.94 (d, 6H, J=6.8 Hz, thexyl), 1.48 (s, 3H, 16-Me),4.85, 5.09 (2s, 2H, ═CH₂), 6.55 (d, 1H, J=2.4 Hz, H-4), 6.61 (dd, 1H,J=2.4 Hz, 8.3 Hz, H-2), 7.13 (d, 1H, J=8.3 Hz, H-1).

Example 16

3,16α-Dihydroxy-16β-methyl-17-methylene-estra-1,3,5(10)-triene

Prepared from3,16α-dihydroxy-16β-methyl-17-methylene-estra-1,3,5(10)-triene,3-O-dimethyl-thexylsilyl ether according to procedure B.

Yield: 56 mg (78%)

R_(f) (1:1)=0.47

mp 238-243° C.

MS(FAB): m/z=298 (M⁺)

¹ H NMR (CDCl₃) δ ¹ H NMR (CDCl₃) δ 0.87 (s, 3H, H-18), 1.49 (s, 3H,16-Me), 4.53 (s, 1H, phenol), 4.86 (s, 1H, ═CH₂), 5.10 (s, 1H, ═CH₂),6.57 (d, 1H, J=2.4 Hz, H-4), 6.64 (dd, 1H, J=2.4 Hz, 8.3 Hz, H-2), 7.18(d, 1H, J=8.3 Hz, H-1).

Example 17

(17Z)-3-Hydroxy-19-Norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether

Potassium tert-butoxide (325 mg, 2.90 mmol) was added to a solution ofethyltriphenylphosphonium bromide (1.08 g, 2.90 mmol) in dry DMSO (6.0ml) under N₂. After stirring for 20 min the temperature was raised to75° C. and a solution of 3-hydroxy-17-keto-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether (1.00 g, 2.42 mmol) in dry THF (4.0 ml)was added. The reaction mixture was stirred at 75° C. for 2.5 h. Aftercooling the reaction mixture was partitioned in Et₂ O/H₂ O and theorganic phase was washed with H₂ O, dried over Na₂ SO₄, and concentratedat reduced pressure. The residue was purified by column chromatography(heptane) to give the title compound as an approximately 1:1 epimericmixture (85 mg, 28%).

R_(f) (heptane)=0.2

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --Si(CH₃)₂ --), 0.91 (s, 3H, H-18), 0.94(s, 6H, thexyl), 0.94 (d, 6H, J=6.8 Hz, thexyl), 1.7 (m, 3H, H-21), 6.54(d, 1H, J=2.6 Hz, H-4), 6.61 (dd, 1H, J=2.6 Hz, 8.7 Hz, H-2), 7.13 (d,1H, J=8.7 Hz, H-1).

Example 18

(17E)-3,16α-Dihydroxy-19-Norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether

Prepared from (17Z)-3-hydroxy-19-norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether according to procedure A.

Yield: 140 mg (51%)

R_(f) (10:1)=0.07

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --Si(CH₃)₂ --), 0.92 (s, 3H, H-18), 0.94(s, 6H, thexyl), 0.94 (d, 6H, J=6.8 Hz, thexyl), 1.78 (d, 3H, J=7 Hz,H-21), 4.48 (s, 1H, H-16), 6.56 (d, 1H, J=2.2 Hz, H-4), 6.62 (dd, 1H,J=2.2 Hz, 8.6 Hz, H-2), 7.12 (d, 1H, J=8.6 Hz, H-1).

Example 19

(17E)-3,16α-Dihydroxy-19-Norpregna-1,3,5(10),17(20)-tetraene

Prepared from(17E)-3,16α-dihydroxy-19-norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethyl-thexylsilyl ether according to procedure B.

Yield: 30 mg (84%)

R_(f) (1:1)=0.39

mp 225-31 ° C.

MS(FAB): m/z=298

¹ H NMR (CDCl₃) δ 0.92 (s, 3H, H-18), 1.78 (d, 3H, J=7 Hz, H-21), 4.48(s, 1H, H-16), 6.57 (d, 1H, J=2.6 Hz, H-4), 6.63 (dd, 1H, J=2.6 Hz, 8.5Hz, H-2), 7.16 (d, 1H, J=8.5 Hz, H-1).

Example 20

Etyl (17E)-3-Hydroxy-19-Norpregna-1,3,5(10),17(20)-tetraene-21-oate,3-O-dimethylthexylsilyl ether

Triethyl phosphonoacetate (3.00 mL, 15.0 mnol) was added dropwise to aslurry of NaH (480 mg, ca 60% in oil, 12 mmol) in dry dimethoxyethane(DME, 30 ml) under N₂. After 10 min stirring, a solution of3-hydroxy-17-keto-estra-1,3,5(10)-triene, 3-dimethylthexylsilyl ether(2.064 g, 5.00 mmol) in dry DME (15 ml) was added. The temperature wasraised to 90° C. and the reaction mixture was then stirred over night.After cooling heptane (20 ml) was added and most of the DME was removedby evaporation at reduced pressure. The residue was partitioned in Et₂O/H₂ O and the organic phase was then washed with brine, dried over Na₂SO₄, and concentrated at reduced pressure. The residue was purified bycolumn chromatography (heptane-EtOAc, 50:1, 20:1) to give the titlecompound as a white solid (1.494 mg, 62%).

R_(f) (20:1)=0.30;

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --Si(CH₃)₂ --), 0.86 (s, 3H, H-18), 0.94(s, 6H, thexyl), 0.94 (d, 6H, J=6.8 Hz, thexyl), 1.29 (t, 3H, J=7.1 Hz,Et), 4.16 (q, 2H, J=7.1 Hz, Et), 5.59 (dd, 1H, J=2.4 Hz, 2.4 Hz, H-20),6.55 (d, 1H, J=2.7 Hz, H-4), 6.61 (dd, 1H, J=2.7 Hz, 8.5 Hz, H-2), 7.12(d, 1H, J=8.5 Hz, H-1).

Example 21

(17E)-3,21-Dihydroxy-19-Norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether

Lithium triethylborohydride (6.0 mL, 1 M in THF, 6.0 mmol) was added toa solution of etyl(17E)-3-hydroxy-19-norpregna-1,3,5(10),17(20)-tetraene-21-oate,3-O-dimethylthexylsilyl ether (1.320 g, 2.73 mmol) in dry THF (6.0 mL)at 0° C. under N₂. The reaction mixture was stirred for another 10 minand was then partitioned in Et₂ O/brine and acidified with 1 M HCl (ca10 mL). The organic phase was washed with brine, dried over Na₂ SO₄, andconcentrated at reduced pressure. The residue was purified by columnchromatography (heptane-EtOAc, 5:1, 3:1) to give the title compound as awhite solid (1.048 mg, 87%).

R_(f) (3:1)=0.27

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --Si(CH₃)₂ --), 0.81 (s, 3H, H-18), 0.94(s, 6H, thexyl), 0.94 (d, 6H, J=6.8 Hz, thexyl), 4.14 (m, 2H, H-21),5.29 (m, 1H, H-20), 6.54 (d, 1H, J=2.4 Hz, H-4), 6.61 (dd, 1H, J=2.4 Hz,8.1 Hz, H-2), 7.13 (d, 1H, J=8.1 Hz, H-1).

Example 22

(17E)-3-Hydroxy-19-Norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether

Methanesulfonic anhydride (52 mg, 0.3 mmol) was added to a solution of(17E)-3,21-dihydroxy-19-norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether (74 mg, 0.17 mmol) and 2,6-lutidine (46μL, 0.4 mmol) in dry THF (0.5 mL) under N₂. After 5 min stirring,lithium triethylborohydride (500 μL, 1 M in THF, 0.50 mmol) was added.The reaction mixture was stirred for another 10 min and was thenpartitioned in Et₂ O/brine and acidified with 1 M HCl (ca 5 mL). Theorganic phase was washed with brine, NaHCO₃ (sat.) and brine again,dried over Na₂ SO₄, and concentrated at reduced pressure. The residuewas purified by column chromatography (heptane-EtOAc, 50:1) to give thetitle compound as an oil (40 mg, 56%).

R_(f) (50:1)=0.30

¹ H NMR (CDCl₃) δ 0.21 (s, 6H, --Si(CH₃)₂ --), 0.77 (s, 3H, H-18), 0.94(s, 6H, thexyl), 0.94 (d, 6H, J=7.1 Hz, thexyl), 1.56 (ddd, 3H, J=6.6Hz, 1.5 Hz, 1.5 Hz, H-21), 5.08 (m, 1H, H-20), 6.54 (d, 1H, J=2.7 Hz,H-4), 6.60 (dd, 1H, J=2.7 Hz, 8.3 Hz, H-2), 7.14 (d, 1H, J=8.3 Hz, H-1).

Example 23

(17Z)-3,16α-Dihydroxy-19-Norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether

Prepared from (17E)-3-hydroxy-19-norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether according to procedure A.

Yield: 25 mg (45%)

R_(f) (5:1)=0.29

¹ H NMR (CDCl₃) δ 0.21 (s, 6H, --Si(CH₃)₂ --), 0.77 (s, 3H, H-18), 0.94(s, 6H, thexyl), 0.94 (d, 6H, J=7 Hz, thexyl), 1.81 (d, 3H, J=7 Hz,H-21), 4.85 (m, 1H, H-16), 5.37 (dq, 1H, J=2 Hz, 7 Hz, H-20), 6.55 (d,1H, J=2.7 Hz, H-4), 6.61 (dd, 1H, J=2.7 Hz, 8.2 Hz, H-2), 7.12 (d, 1H,J=8.2 Hz, H-1).

Example 24

(17Z)-3-Hydroxy-16-keto-19-Norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethylthexylsilyl ether

The reaction according to Example 23 also provided the compound of thisExample.

Yield: 20 mg (36%)

R_(f) (10:1)=0.19

¹ H NMR (CDCl₃) δ 0.22 (s, 6H, --Si(CH₃)₂ --), 0.94 (s, 6H, thexyl),0.94 (d, 6H, J=6.8 Hz, thexyl), 1.06 (s, 3H, H-18), 1.89 (d, 3H, J=7.6Hz, H-21), 6.54 (q, 1H, J=7.6 Hz, H-20), 6.56 (d, 1H, J=2.7 Hz, H-4),6.63 (dd, 1H, J=2.7 Hz, 8.5 Hz, H-2), 7.12 (d, 1H, J=8.5 Hz, H-1).

Example 25

(17Z)-3,16α-Dihydroxy-19-Norpregna-1,3,5(10),17(20)-tetraene

Prepared from(17Z)-3,16α-dihydroxy-19-norpregna-1,3,5(10),17(20)-tetraene,3-O-dimethyl-thexylsilyl ether according to procedure B.

Yield: 11 mg (82%)

R_(f) (2:1)=0.26

mp 228-32° C.

MS-FAB: m/z=298 (M⁺)

¹ H NMR (CDCl₃) δ 0.77 (s, 3H, H-18), 1.81 (d, 3H, J=6.8 Hz, H-21), 4.57(s, 1H, 3-OH), 4.85 (m, 1H, H-16), 5.38 (dq, 1H, J=1.7 Hz, 6.8 Hz,H-20), 6.57 (d, 1H, J=2.7 Hz, H-4), 6.63 (dd, 1H, J=2.7 Hz, 8.5 Hz,H-2), 7.16 (d, 1H, J=8.5 Hz, H-1).

Example 26

3,16α,17β-Trihydroxy-estra-1,3,5(10)-triene,3,16α-Bis(dimethylthexylsilyl ether)

Dimethylthexylchlorosilane (1.47 ml, 7.49 mmol) was added to a solutionof 3,16α,17β-trihydroxy-estra-1,3,5(10)-triene (estriol, 1.00 g, 3.47mmol) and imidazole (1.02 g, 15.0 mmol) in dry DMF (2.0 ml). Thereaction mixture was stirred for 30 min and the raw product was thenpurified directly by column chromatography (heptane-EtOAc, 10:1) to givethe title compound as an oil which crystallized on standing (1.95 g,98%).

R_(f) (10:1)=0.22

¹ H NMR (CDCl₃) δ 0.11 (s, 3H, --SiMe₂ --), 0.13 (s, 3H, --SiMe₂ --),0.21 (s, 6H, --SiMe₂ --), 0.78 (s, 3H, H-18), 0.86 (s, 6H, thexyl), 0.90(d, 6H, J=6.8 Hz, thexyl), 0.94 (s, 6H, thexyl), 0.94 (d, 6H, J=6.8 Hz,thexyl), 3.56 (t, 1H, J=5.4 Hz, H-17), 4.07 (m, 1H, H-16), 6.54 (d, 1H,J=2.7Hz, H-4), 6.60 (dd, 1H, J=2.7 Hz, 8.3Hz, H-2), 7.11 (d, 1H, J=8.3Hz, H-1).

Example 27

3,16α-Dihydroxy-17-keto-estra-1,3,5(10)-triene,3,16α-Bis(dimethyl-thexylsilyl ether)

N-methylmorphonlin (300 mg, 2.22 mmol) and tetrapropylammoniumperruthenate (TPAP, 40 mg, 0.11 mmol) were added to a solution of3,16α,17β-trihydroxy-estra-1,3,5(10)-triene,3,16α-bis(dimethyl-thexylsilyl ether) (790 mg, 1.38 mmol) in CH₂ Cl₂(3.0 ml). The solution was stirred for 6 h at room temperature and wasthen concentrated at reduced pressure. The residue was purified bycolumn chromatography (heptane-EtOAc, 50:1, 20:1) to give the titlecompound as an oil (600 mg, 76%).

R_(f) (20:1)=0.33

¹ H NMR (CDCl₃) δ 0.15 (s, 3H, --SiMe₂ --), 0.18 (s, 3H, --SiMe₂ --),0.22 (s, 6H, --SiMe₂ --), 0.86 (s, 6H, thexyl), 0.88 (d, 3H, J=6.8 Hz,thexyl), 0.89 (d, 3H, J=6.8 Hz, thexyl), 0.93 (s, 3H, H-18), 0.94 (s,6H, thexyl), 0.94 (d, 6H, J=6.8 Hz, thexyl), 4.36 (d, 1H, J=7.5 Hz,H-16), 6.55 (d, 1H, J=2.7 Hz, H-4), 6.61 (dd, 1H, J=2.7 Hz, 8.3 Hz,H-2), 7.11 (d, 1H, J=8.3 Hz, H-1).

Example 28

17-Difluoromethylene-3,16α-dihydroxy-estra-1,3,5(10)-triene,3,16α-Bis(dimethylthexylsilyl ether)

Lithium diisopropylamide (750 μl, 1.5 M THF-complex in hexane, 1.12mmol) was added to a solution of F₂ CHPO(OEt)₂ (215 mg, 1.14 mmol) indry THF (1.0 ml) under N₂ at -78° C. After 5 min stirring, a solution of3,16α-dihydroxy-17-keto-estra-1,3,5(10)-triene,3,16α-bis(dimethyl-thexylsilyl ether) (173 mg, 0.30 mmol) in dry THF wasadded and the reaction mixture was stirred at -78° C. for 1 h, then at60° C. over night. After cooling, the reaction mixture was diluted withEt₂ O (100 ml) and acidified with 1M HCl. The organic phase was washedwith brine, dried over Na₂ SO₄ and concentrated at reduced pressure. Theresidue (268 mg of a brown oil) was purified by column chromatography(heptane, then heptane-EtOAc, 50:1, then 20:1) to give the titlecompound as an oil (102 mg, 56%).

R_(f) (50:1)=0.33

¹ H-NMR (CDCl₃) δ 0.11 (s, 6H, --SiMe₂ --), 0.21 (s, 6H, --SiMe₂ --),0.82 (s, 6H, thexyl), 0.87 (2d, 6H, J=6 Hz, thexyl), 0.88 (s, 3H, H-18),0.94 (s, 6H, thexyl), 0.94 (d, 6H, J=6.9 Hz, thexyl), 4.77 (dd, 1H,J=1.6 Hz, 5.2 Hz, H-16), 6.54 (d, 1H, J=2.7 Hz, H-4), 6.61 (dd, 1H,J=2.7 Hz, 8.4 Hz, H-2), 7.11 (d, 1H, J=8.4 Hz, H-1).

Example 29

17-Difluoromethylene-3,16α-dihydroxy-estra-1,3,5(10)-triene

NBu₄ F.(H₂ O)₃ (200 mg, 0.63 mmol) was added to a solution of17-difluoromethylene-3,16α-dihydroxy-estra-1,3,5(10)-triene,3,16α-bis(dimethyl-thexylsilyl ether) (100 mg, 0.165 mmol) in dry THF(1.0 ml). The reaction mixture was stirred for 2 h at 50° C. and wasthen quenched by adding AcOH (100 μl). Concentration at reduced pressurewas followed by purification by column chromatography (heptane-EtOAc,3:1, 2:1) to give the title compound as a white solid (19 mg, 36%):

R_(f) (2:1)=0.28

mp 225-27° C.

MS-FAB: m/z=320 (M⁺)

¹ H NMR (CDCl₃) δ 0.91 (s, 3H, H-18), 4.52 (s, 1H, 3-OH), 4.89 (m, 1H,H-16), 6.57 (d, 1H, J=2.7 Hz, H-4), 6.63 (dd, 1H, J=2.7 Hz, 8.5 Hz,H-2), 7.15 (d, 1H, J=8.5 Hz, H-1).

3-O-Alkylether derivatives

General Procedure for 3-O-alkylation of3,16α-dihydroxy-17-methylene-estra-1,3,5(10)-triene

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene (0.32 mmol), alkyliodide (0.42 mmol), Cs₂ CO₃ (0.70 mmol) and dry DMF (0.5-1.0 mL) underdry nitrogen were stirred over night at 40-80° C. The volatiles wereevaporated at reduced pressure and the residue was partitioned betweensaturated NH₄ Cl and EtOAc (2×10 mL). The organic phases were combined,washed with brine, dried over anhydrous Na₂ SO₄, filtered andconcentrated at reduced pressure. The residue was purified by columnchromatography on silica (heptane-EtOAc, 5:1) to give the3-O-alkylether.

The following 3-O-alkyl ethers were prepared:

Example 30

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-O-Cyclopentylether

Yield: 59%; colourless crystalline solid

R_(f) (3:1)=0.25

MS(EI) m/z 352 (M⁺)

¹ H NMR (CDCl₃) δ 0.82 (s, 3H, H-18), 4.67-4.75 (m, 2H), 4.92 (d, 1H,J=1.8 Hz, ═CH₂), 5.08 (d, 1H, J=1.5 Hz, ═CH₂), 6.60 (d, 1H, J=2.6 Hz,H-4), 6.67 (dd, 1H, J=8.4 Hz, J=2.6 Hz, H-2), 7.18 (d, 1H, J=8.4 Hz,H-1).

Example 31

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-O-methyl ether

Yield: 49%; colourless crystalline solid

R_(f) (2:1)=0.24

MS(EI) m/z 298 (M⁺)

¹ H NMR (CDCl₃) δ 0.83 (s, 3H, H-18), 3.78 (s, 3H, --OCH₃), 4.69-4.75(m, 1H, H-16), 4.93 (d, 1H, J=2.4 Hz, ═CH₂), 5.09 (d, 1H, J=1.8 Hz,═CH₂), 6.64 (d, 1H, J=2.7 Hz, H-4), 6.72 (dd, 1H, J=8.4 Hz, J=2.7 Hz,H-2), 7.22 (d, 1H, J=8.7 Hz, H-1).

Ester and carbonic-acid ester derivatives

General Procedure for 3-O-Monoesterification of3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene:

An acid chloride or chloroformate ester (0.36 mmol) in dry dioxane (0.35mL) was added during 15 minutes to a rapidly stirred mixture of3,16α-dihydroxy-17-methylene-estra-1,3,5(10)-triene (0.090 g, 0.32mmol), ground NaOH (0.035 g), tetrabutylammonium hydrogen sulfate (2-4mg) and dioxane (0.80 mL). After stirring at room temperature for 10-30minutes saturated NH₄ Cl (2 mL), water (0.5 mL) and EtOAc (10 mL) wereadded. The organic phase was separated, washed with brine, dried overanhydrous Na₂ SO₄, filtered and concentrated at reduced pressure. Theresidue was purified by column chromatography on silica (with the eluentindicated below) to give the title compound.

Yields: 40-60%.

The following 3-O-monoesters were prepared:

Example 32

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-acetate

R_(f) (1:1)=0.31

¹ H NMR (CDCl₃) δ 0.83 (s, 3H, H-18), 2.28 (s, 3H, Ac), 4.72 (m, 1H,H-16), 4.93 (d, 1H, J=2.2 Hz, ═CH₂), 5.09 (d, 1H, J=1.7 Hz, ═CH₂), 6.80(d, 1H, J=2.4 Hz, H-4), 6.85 (dd, 1H, J=2.4 Hz, 8.6 Hz, H-2), 7.29 (d,1H, J=8.6 Hz, H-1).

Example 33

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-Benzoate

R_(f) (3:1)=0.20

¹ H NMR (CDCl₃) δ 0.85 (s, 3H, H-18), 4.73 (m, 1H, H-16), 4.94 (d, 1H,J=2.0 Hz, ═CH₂), 5.10 (d, 1H, J=1.7 Hz, ═CH₂), 6.93 (d, 1H, J=2.4 Hz,H-4), 6.98 (dd, 1H, J=2.4 Hz, 8.3 Hz, H-2), 7.35 (d, 1H, J=8.3 Hz, H-1),7.51 (m, 2H, Bz), 7.63 (m, 1H, Bz), 8.20 (m, 2H, Bz).

Example 34

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-Hexanoate

R_(f) (1:1)=0.37

¹ H NMR (CDCl₃) δ 0.84 (s, 3H, H-18), 0.90-0.98 (m, 3H), 2.54 (t, J=7.5Hz, 2H), 4.69-4.76 (m, 1H, H-16), 4.91 (d, 1H, J=2.4 Hz, ═CH₂), 5.10 (d,1H, J=1.8 Hz, ═CH₂), 6.80 (d, 1H, J=2.4 Hz, H-4), 6.85 (dd, 1H, J=8.4Hz, J=2.4 Hz, H-2), 7.30 (d, 1H, J=8.4 Hz, H-1).

Example 35

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-Octadecanoate

R_(f) (2:1)=0.29

¹ H NMR (CDCl₃) δ 0.83 (s, 3H, H-18), 0.85-0.92 (m, 3H), 2.53 (t, J=7.5Hz, 2H), 4.68-4.76 (m, 1H, H-16), 4.92-4.94 ("d", 1H, ═CH₂), 5.07-5.11("d", 1H, ═CH₂), 6.77-6.80 (m, 1H, H-4), 6.81-6.86 (m, 1H, H-2) and 7.28(d, 1H, J=9.0 Hz, H-1).

Example 36

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-methylcarbonate

R_(f) (2:1)=0.19

¹ H NMR (CDCl₃) δ 0.83 (s, 3H, H-18), 3.89 (s, 3H, --OCH₃), 4.68-4.75(m, 1H, H-16), 4.93 (d, 1H, J=2.1 Hz, ═CH₂), 5.09 (d, 1H, J=1.8 Hz,═CH₂), 6.88 (d, 1H, J=2.4 Hz, H-4), 6.93 (dd, 1H, J=8.4 Hz, J=2.4 Hz,H-2), 7.29 (d, 1H, J=8.4 Hz, H-1).

Example 37

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-Butylcarbonate

R_(f) (1:1)=0.45

¹ H NMR (CDCl₃) δ 0.83 (s, 3H, H-18), 0.97 (t, J=7.2 Hz, 3H), 4.24 (t,J=6.6 Hz, 2H), 4.68-4.75 (m, 1H, H-16), 4.92 (d, 1H, J=1.8 Hz, ═CH₂),5.08 (d, 1H, J=1.2 Hz, ═CH₂), 6.89 (d, 1H, J=2.4 Hz, H-4), 6.94 (dd, 1H,J=8.4 Hz, J=2.4 Hz, H-2) and 7.29 (d, 1H, J=8.1 Hz, H-1).

Example 38

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3-Benzylcarbonate

R_(f) (2:1)=0.21

¹ H NMR (CDCl₃) δ 0.82 (s, 3H, H-18), 4.68-4.74 (m, 1H, H-16), 4.92 (d,1H, J=1.8 Hz, ═CH₂), 5.08 (d, 1H, J=1.5 Hz, ═CH₂), 5.25 (s, 2H, OCH₂Ph), 6.88 (d, 1H, J=2.4 Hz, H-4), 6.94 (dd, 1H, J=8.7 Hz, J=2.7 Hz,H-2), 7.29 (d, 1H, J=8.4 Hz, H-1), 7.34-7.46 (m, 5H, C₆ H₅ --).

General Procedure for 16α-O-Monoesterification of3,16α-dihydroxy-17-methylene-estra-1,3,5(10)-triene:

An ester anhydride or ester chloride (1.1 mmol) was added to a solutionof 3,16α-dihydroxy-D17-methylene-estra-1,3,5(10)-triene,3-O-dimethylthexylsilyl ether (1.0 mmol) and N,N-dimethylaminopyridine(1.5 mmol) in CH₂ Cl₂ (1.5 mL). The reaction mixture was stuffed for 1-4h and was then concentrated at reduced pressure. The residue wasfiltered through a short silica gel column (heptane-EtOAc mixtures aseluents). The filtrate was concentrated at reduced pressure and theresidue was treated according to Procedure B.

Yields: 60-80%.

The following 16α-O-monoester derivatives were prepared:

Example 39

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 16α-Acetate

R_(f) (5:1)=0.17

¹ H NMR (CDCl₃) δ 0.85 (s, 3H, H-18), 2.11 (s, 3H, Ac), 4.64 (s, 1H,phenol), 4.94 (d, 1H, J=2.0 Hz, ═CH₂), 4.97 (d, 1H, J=1.7 Hz, ═CH₂),5.72 (broad d, 1H, J=7.8 Hz, H-16), 6.57 (d, 1H, J=2.7 Hz, H-4), 6.63(dd, 1H, J=2.7 Hz, 8.6 Hz, H-2), 7.16 (d, 1H, J=8.6 Hz, H-1).

Example40

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 16α-Hexanoate

R_(f) (5:1)=0.22

¹ H NMR (CDCl₃) δ 0.84 (s, 3H, H-18), 0.90 (t, 3H, J=7 Hz), 2.35 (t, 2H,J=7.5 Hz), 4.93 (d, 1H, J=2.1, Hz, ═CH₂), 4.95 (d, 1H, J=1.8 Hz, ═CH₂),5.73 (d, 1H, J=6.9 Hz, H-16), 6.57 (d, 1H, J=2.7 Hz, H-4), 6.65 (dd, 1H,J=2.7 Hz, 8.4 Hz, H-2), 7.15 (d, 1H, J=8.4 Hz, H-1).

Example 41

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 16α-Octadecanoate

R_(f) (5:1)=0.23

¹ H NMR (CDCl₃) δ 0.86 (s, 3H, H-18), 0.88 (t, 3H, J=7 Hz), 2.34 (t, 2H,J=7.5 Hz), 4.55 (s, 1H, phenol), 4.93 (d, 1H, J=2.3, Hz, ═CH₂), 4.95 (d,1H, J=1.7 Hz, ═CH₂), 5.73 (broad d, 1H, J=6.3 Hz, H-16), 6.56 (d, 1H,J=2.9 Hz, H-4), 6.64 (dd, 1H, J=2.9 Hz, 8.2 Hz, H-2), 7.16 (d, 1H, J=8.2Hz, H-1).

Example 42

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 16α-Benzoate

R_(f) (5:1)=0.12

¹ H NMR (CDCl₃) δ 0.92 (s, 3H, H-18), 4.53 (s, 1H, phenol), 4.99 (d, 1H,J=2.1, Hz, ═CH₂), 5.07 (d, 1H, J=1.8 Hz, ═CH₂), 5.95 (broad d, 1H, J=6.6Hz, H-16), 6.57 (d, 1H, J=2.7 Hz, H-4), 6.64 (dd, 1H, J=2.7 Hz, 8.7 Hz,H-2), 7.18 (d, 1H, J=8.7 Hz, H-1), 7.45 (t, 2H, J=7.4 Hz, Ph), 7.57 (m,1H, Ph), 8.08 (d, 2H, J=7.4 Hz, Ph).

Example 43

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 16α-methylcarbonate

R_(f) (3:1)=0.27

¹ H NMR (CDCl₃) δ 0.85 (s, 3H, H-18), 3.81 (s, 3H, OMe), 4.76 (s, 1H,phenol), 5.00 (d, 1H, J=2.2 Hz, ═CH₂), 5.05 (d, 1H, J=1.7 Hz, ═CH₂),5.59 (m, 1H, H-16), 6.57 (d, 1H, J=2.7 Hz, H-4), 6.63 (dd, 1H, J=2.7 Hz,8.3 Hz, H-2), 7.16 (d, 1H, J=8.3 Hz, H-1).

Example 44

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene,16α-n-Butylcarbonate

R_(f) (5:1)=0.16

¹ H NMR (CDCl₃) δ 0.85 (s, 1H, H-18), 0.94 (t, J=7.5 Hz, 3H), 4.17 (t,2H, J=6.6 Hz), 4.99 (s, 1H, ═CH₂), 5.10 (s, 1H, ═CH₂), 5.60 (m, 1H,H-16), 6.57 (d, 1H, J=2.7 Hz, H-4), 6.62 (dd, 1H, J=2.7 Hz, 8.5 Hz,H-2), 7.16 (d, 1H, J=8.5 Hz, H-1).

Example 45

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 16α-Benzylcarbonate

R_(f) (5:1)=0.14

¹ H NMR (CDCl₃) δ 0.85 (s, 1H, H-18), 4.98 (d, 1H, J=1.8 Hz, ═CH₂), 5.09(d, 1H, J=1.5 Hz, ═CH₂), 5.19 (s, 2H, benzyl), 5.62 (m, 1H, H-16), 6.56(d, 1H, J=2.7 Hz, H-4), 6.63 (dd, 1H, J=2.7 Hz, 8.4 Hz, H-2), 7.15 (d,1H, J=8.4 Hz, H-1), 7.33-7.42 (m, 5H, Ph).

General Procedure for 3-O,16α-O-diesterification of3,16α-dihydroxy-17-methylene-estra-1,3,5(10)-triene:

An ester anhydride or ester chloride (3.0 mmol) was added to a solutionof 3,16α-dihydroxy-17-methylene-estra-1,3,5(10)-triene (1.0 mmol) andN,N-dimethyl-aminopyridine (4.0 mmol) in CH₂ Cl₂ (1.5 nmL). The reactionmixture was stirred for 1-3 h and was then concentrated at reducedpressure. The residue was purified by column chromatography(heptane-EtOAc) to give the 3,16α-diester derivatives.

Yields ca 70-80%.

The following 3-O,16α-O-diesters were prepared:

Example 46

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3,16α-diacetate

R_(f) (10:1)=0.33

¹ H NMR (CDCl₃) δ 0.85 (s, 3H, H-18), 2.10 (s, 3H, Ac), 2.28 (s, 3H,Ac), 4.95 (d, 1H, J=2.0 Hz, ═CH₂), 4.98 (d, 1H, J=1.5 Hz, ═CH₂), 5.73(broad d, 1H, J=7.8 Hz, H-16), 6.80 (d, 1H, J=2.4 Hz, H-4), 6.85 (dd,1H, J=2.4 Hz, 8.6 Hz, H-2), 7.29 (d, 1H, J=8.6 Hz, H-1).

Example 47

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3,16α-dihexanoate

R_(f) (5:1)=0.61

¹ H NMR (CDCl₃) δ 0.86 (s, 1H, H-18), 0.91 (m, 6H), 2.34 (t, 2H, J=7.5Hz), 2.53 (t, 2H, J=7.5 Hz), 4.93 (d, 1H, J=2.2 Hz, ═CH₂), 4.95 (d, 1H,J=1.7 Hz, ═CH₂), 5.73 (broad d, 1H, J=6.9 Hz, H-16), 6.78 (d, 1H, J=2.5Hz, H-4), 6.84 (dd, 1H, J=2.5 Hz, 8.4 Hz, H-2), 7.29 (d, 1H, J=8.4 Hz,H-1).

Example 48

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene,3,16α-dihexadecanoate

R_(f) (20:1)=0.27

¹ H NMR (CDCl₃) δ 0.84-0.92 (m, 9H), 2.34 (t, 2H, J=7.5 Hz), 2.53 (t,2H, J=7.5 Hz), 4.93 (d, 1H, J=2.2 Hz, ═CH₂), 4.95 (d, 1H, J=1.7 Hz,═CH₂), 5.73 (broad d, 1H, J=7.6 Hz, H-16), 6.78 (d, 1H, J=2.4 Hz, H-4),6.84 (dd, 1H, J=2.4 Hz, 8.5 Hz, H-2), 7.29 (d, 1H, J=8.5 Hz, H-1).

Example 49

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene, 3,16α-dibenzoate

R_(f) (5:1)=0.34

¹ H NMR (CDCl₃) δ 0.93 (s, 1H, H-18), 5.00 (d, 1H, J=2 Hz, ═CH₂), 5.08(d, 1H, J=2 Hz, ═CH₂), 5.96 (d, 1H, J=7.5 Hz, H-16), 6.94 (d, 1H, J=2.4Hz, H-4), 6.99 (dd, 1H, J=2.4 Hz, 8.7 Hz, H-2), 7.46 (d, 1H, J=8.7 Hz,H-1), 7.30-7.70 (m, 4H), 8.07-8.22 (m, 6H).

Example 50

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene,3,16α-di(methylcarbonate)

R_(f) (5:1)=0.24

¹ H NMR (CDCl₃) δ 0.85 (s, 1H, H-18), 3.81 (s, 3H, --OCH₃), 3.89 (s, 3H,--OCH₃), 5.03 (d, 1H, J=2.2 Hz, ═CH₂), 5.10 (d, 1H, J=1.7 Hz, ═CH₂),5.59 (m, 1H, H-16), 6.89 (d, 1H, J=2.4 Hz, H-4), 6.93 (dd, 1H, J=2.4 Hz,8.4 Hz, H-2), 7.29 (d, 1H, J=8.4 Hz, H-1).

Example 51

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene,3,16α-di(n-butylcarbonate)

R_(f) (5:1)=0.50

¹ H NMR (CDCl₃) δ 0.85 (s, 1H, H-18), 0.96 (m, 6H), 4.17 (t, 2H, J=6.8Hz), 4.24 (t, 2H, J=6.8 Hz), 4.99 (d, 1H, J=2.1 Hz, ═CH₂), 5.11 (d, 1H,J=1.8 Hz, ═CH₂), 5.60 (m, 1H, H-16), 6.89 (d, 1H, J=2.4 Hz, H-4), 6.93(dd, 1H, J=2.4 Hz, 8.5 Hz, H-2), 7.28 (d, 1H, J=8.5 Hz, H-1).

Example 52

3,16α-Dihydroxy-17-methylene-estra-1,3,5(10)-triene,3,16α-di(benzylcarbonate)

R_(f) (5:1)=0.23

¹ H NMR (CDCl₃) δ 0.84 (s, 1H, H-18), 4.99 (d, 1H, J=2 Hz, ═CH₂), 5.10(d, 1H, J=2 Hz, ═CH₂), 5.19, 5.26 (2s, 4H, benzyl), 5.62 (m, 1H, H-16),6.88 (d, 1H, J=2.4 Hz, H-4), 6.94 (dd, 1H, J=2.4 Hz, 8.4 Hz, H-2),7.26-7.43 (m, 11H, H-1, Ph).

Example 53

17-(1',2'-Ethylene)-3-hydroxy-16-keto-estra-1,3,5(10)-trienene,3-dimethyl-thexylsilyl ether

NaH (55-65% in oil, 120 mg, 3.0 mmol) was washed under N₂ three timeswith dry n-hexane and dried at reduced pressure. Dry DMSO (3.0 mL) wasthen added followed by finely ground and vacuum-driedtrimethylsulfoxonium iodide (662 mg, 3.0 mmol). The mixture was stirredunder nitrogen until the hydrogen gas evolution ceased and the solutionbecame clear (within 20 min), then transferred dropwise to a stirredsolution of 3-hydroxy-16-keto-17-metylene-estra-1,3,5(10)-trienene,3-dimethyl-thexylsilyl ether (1.27 g, 3.0 mmol) in dry DMSO (2.0 mL) anddry THF (2.0 mL. After stirring for 2 h at room temperature EtOAc (20mL) was added and the solution was washed five times with 5% aqueousNaCl. Then the organic phase was washed with brine, dried over anhydrousNa₂ SO₄, filtered and concentrated at reduced pressure. The residualyellow oil was purified by column chromatography (toluene as eluent) togive the title compound (230 mg, 18%) as a colourless oil, whichsolidified upon cooling.

TLC: R_(f) (toluene)=0.21

MS(EI) m/z 438 (M⁺)

¹ H NMR (CDCl₃) δ 0.23 (s, 6H), 0.65-0.70 (m, 1H), 0.76-0.81 (m, 1H),0.95 (s, 3H), 0.96 (s, 6H), 0.97 (d, 6H), 1.02-1.07 (m, 1H), 1.19-1.24(m, 1H), 1.36-1.40 (m, 2H), 1.44-1.52 (m, 1H), 1.56-1.78 (m, 3H),1.83-1.92 (m, 2H), 2.21 (app dd, 1H, J=14 Hz, J=17 Hz, 2.35-2.44 (m,3H), 2.81-2.93 (m, 2H), 6.58 (d, 1II, J=2.4 Hz), 6.63 (dd, 1H, J=8.3 Hz,J=2.7 Hz), 7.13 (d, 1H, J=8.5 Hz).

Pharmaceutical Preparations

The novel steroidal estrogens according to the invention may beadministered by transdermal patches, orally or intranasally.

The dosage will depend on the route of administration, the severity ofthe disease, age and weight of the patient and other factors normallyconsidered by the attending physician, when determining the individualregimen and dosage level as the most appropriate for a particularpatient.

The pharmaceutical preparation comprising a compound of the inventionmay be a patch, a tablet, a capsule or a nasal spray.

In a transdermal device, an estrogen compound of the invention isdissolved in suitable solvents (e.g. ethanol, propylene glycol)comprising a thickener. The patch further comprises a current backingmembrane and a silicone release liner. The device may also beconstructed with a rate control membrane.

When administered orally the estrogen compounds of the invention may beadministered as a conventional tablet or gelatin capsule. The tablet maycomprise usual tablet constituents, e.g. diluents (such as lactose),binders (such as polyvidone), lubricants (such as magnesium stearate)and disintegrants (such as microcrystalline cellulose). The estrogencompound may also be mixed with diluents and filled into gelatincapsules.

When administered intranasally by means of a nasal spray, theformulation is a suspension of the novel estrogens of the invention inwater comprising a thickener, a surface active ingredient and apreservative.

Biological Evaluation

The anti-inflammatory and immunosuppressive potencies were evaluated inanimal models for autoimmune diseases.

For rheumatoid arthritis the type II collagen induced arthritis (CIA)model in mice was used (Jansson, L., Holmdahl, R., Clin. Exp. Immunol.(1992), 89, 446-451).

Mouse CIA Model

In this model F1 generation (females) between B10Q and DBA/1 mice areused. The mice are ovariectomized two weeks before induction ofarthritis.

Immunisation is performed using collagen type II (purified from ratchondrosarcoma) emulsified in Freunds complete adjuvant.

The treatanent is performed by subcutaneous administration of estrogenanalogues (0.1 ml) in Miglyol oil vehicle or solutol. The mice aretreated on day 14, 17, 21, 24, 28, and 32 respectively, afterimmunisation. Day 36 is the end of the experiment, and the arthritissymptoms start approximately on day 14-20.

Evaluation of sex-related effects is performed by observing the stage ofestrus by vaginal smears 17, 21, 24, 30, and 36 days after immunisation.At day 36 which is the end of the experiment, the weight of the uterusis recorded.

The evaluation of the arthritic effect is performed by observing thejoints of the paws and legs for swelling and erythema every third dayafter immunisation.

The development of arthritis was evaluated continuously for each groupas the incidence (%) of affected animals. The cumulative incidence (areaunder the curve, "auc") was calculated in each group up to day 30. Theanti-arthritic effect of estrogen treatment was expressed as the auc oftreated animals relative the auc of the control group (auc_(treated)animals /auc_(control), %), i.e. 100% denotes no anti-arthritic effectand 0% denotes total blockade of arthritic development. Theantiarthritic effect is related in dose-response studies to the extentof uterine proliferation, and it is possible to estimate the differencein immunosuppressive/sex hormonal profiles.

The novel steroidal estrogens of the present invention, derivatives of17-alkylidene-3,16-dihydroxy-estra-1,3,5(10)-trienes, show very low "sexhormonal" activity while retaining their anti-inflammatory andimmunosuppressive effects.

The Rat-CIA Model

Still another animal model for the evaluation of the anti-inflammatoryand immunosuppressive effects is the rat CIA model.

In this model female rats of the Dark Agouti strain are used. The ratsare ovariectomized two weeks before induction of arthritis.

Immunisation is performed using the same protocols as for CIA in mice,but with Freunds incomplete adjuvant.

Evaluation of the arthritic and sex-related effects are the same as inthe mouse model. The length of the rat CIA-experiment is 21 days.

We claim:
 1. A compound according to the formula I ##STR7## wherein A ishydrogen, C₂ -C₁₈ alkanoyl, (C₆ aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl,or (C₆ aryloxy)carbonyl, or a protecting group;B is hydrogen, methyl, orethyl; R is hydrogen, a straight, branched or cyclic C₁ -C₆ alkyl, C₂-C₁₈ alkanoyl, (C₆ aryl)carbonyl, C₂ -C₁₉ alkoxycarbonyl, (C₆aryloxy)carbonyl, or a protecting group; X¹ is hydrogen, methyl, ethylor halogen; X² is hydrogen, methyl, ethyl or halogen; and Y is methyleneor a single bond; and pharmaceutically acceptable salts thereof; thecompounds(17E)-16α-Acetoxy-3-methoxy-19-norpregna-1,3,5(10),17(20)-tetraene;(17E)-16α-Hydroxy-3-methoxy-19-norpregna-1,3,5(10),17(20)-tetraene; and(17E)-16β-Hydroxy-3-methoxy-19-norpregna-1,3,5(10),17(20)-tetraenebeingexcluded.
 2. A compound according to claim 1, whereinA is hydrogen orC₂₋₆ alkanoyl; B is hydrogen or methyl; R is hydrogen, a straight,branched or cyclic C₁₋₆ alkyl, C₂₋₁₈ alkanoyl, (C₆ aryl)carbonyl, C₂₋₁₉alkoxycarbaonyl, (C₆ aryloxy)carbonyl, or a protecting group; X¹ ishydrogen, methyl, or fluorine; X² is hydrogen, methyl, or fluorine; andY is a methylene group or a single bond.
 3. A compound according toclaim 1, whereinA is hydrogen or C₂₋₆ alkanoyl; B is hydrogen; R ishydrogen, a straight, branched or cyclic C₁₋₆ alkyl, C₂₋₁₈ alkanoyl, (C₆aryl)carbonyl, C₂₋₁₉ alkoxycarbonyl, (C₆ aryloxy)carbonyl, or aprotecting group; X¹ is hydrogen or fluorine; X² is hydrogen orfluorine; and Y is a single bond or a methylene group.
 4. A compoundaccording to claim 1, whereinA is hydrogen; B is hydrogen; R is hydrogenor C₂ -C₆ alkanoyl; X¹ is hydrogen; X² is hydrogen; Y is a single bond;and the 16-OH group is in the α-position.
 5. A compound of claim 1,having the formula 3,16α-dihydroxy-17-methylene-estra-1,3,5(10)triene.6. A therapeutic composition comprising the compound of claim 1 in apharmaceutically acceptable carrier.
 7. A method for treating rheumatoidarthritis or multiple sclerosis comprising administering to a patient inneed thereof an affective amount of the compound of claim
 1. 8. Themethod of claim 7, whereinB is hydrogen; X¹ is hydrogen or fluorine; andX² is hydrogen or fluorine.
 9. The method of claim 7, whereinA ishydrogen; B is hydrogen; R is hydrogen or C₂₋₆ alkanoyl; X¹ is hydrogen;X² is hydrogen; Y is a single bond; and the 16-OH group is in theα-position.
 10. The method of claim 7, wherein the compound has theformula 3,16α-dihydroxy-17-methylene-estra-1,3,5(10)triene.
 11. Themethod of claim 7, whereinA is hydrogen or C₂ -C₆ alkanoyl; B ishydrogen or methyl; X¹ is hydrogen, methyl, or fluorine; and X₂ ishydrogen, methyl, or fluorine.
 12. A method for treating multiplesclerosis comprising administering to a patient in need thereof aneffective amount of the compound of claim
 1. 13. The method of claim 12,whereinA is hydrogen or C₂ -C₆ alkanoyl; B is hydrogen or methyl; X¹ ishydrogen, methyl, or fluorine; and X² is hydrogen, methyl, or fluorine.14. The method of claim 12, whereinA is hydrogen or C₂ -C₆ alkanoyl; Bis hydrogen; X¹ is hydrogen or fluorine; and X² is hydrogen or fluorine.15. The method of claim 12, whereinA is hydrogen; B is hydrogen; R ishydrogen or C₂ -C₆ alkanoyl; X¹ is hydrogen; X² is hydrogen; Y is asingle bond; and the 16-OH group is in the α-position.
 16. The method ofclaim 12, wherein the compound has the formula3,16α-dihydroxy-17-methylene-estra-1,3,5(10)triene.